湖南省日本血吸虫线粒体atp6基因部分序列的多态性分析

DNA polymorphism in the partial mitochondrial atp6 gene among Schistosoma japonicum isolates from Hunan Province

目的研究湖南省日本血吸虫不同地域自然隔离群线粒体ATP合成酶F0亚单位6基因(atp6)部分序列的遗传多态性,为湖南不同地域自然隔离群日本血吸虫的遗传特性研究提供实验依据。方法采用试剂盒抽提基因组总DNA,以特异性引物对线粒体atp6基因进行PCR扩增,通过单链构象多态性技术(SSCP)筛选出差异带型并进行测序,用DNAStar 5.0及Mega 4.0软件进行比对分析。结果湖南省5个流行区的日本血吸虫PCR扩增后获得了483 bpatp6部分序列,检测出17个变异位点,变异率为3.52%,雌雄虫之间的差异为0.0%～1.3%,不同地理来源虫株间的差异率为0.0%～1.5%。聚类分析结果表明:湖南不同地域自然隔离群日本血吸虫的种系发育不能按照其来源归为相应的系统发育进化树。结论湖南不同地域自然隔离群日本血吸虫的atp6基因存在明显的个体差异,是否由于生存环境的选择压力造成了同一地理来源日本血吸虫个体之间atp6基因的遗传差异,其原因尚有待于进一步研究。

The aim of this study was to examine sequence variation in the partial mitochondrial ATP synthase F0 subunit atp6 gene of Schistosoma japonicum and to provide foundation for studying the characterization and genetic structure of S.japonicum from different endemic origins of Hunan province.After extracting genomic DNA,atp6 was amplified with specific primers and screened by SSCP technique.Representative samples with variable banding patterns were selected for sequencing.DNAStar 5.0 and Mega 4.0 were used to analyze the sequences subsequently.Results showed that partial atp6 gene of S.japonicum was 483 bp,and 17 mutation sites were detected（3.52%）.Sequence variations for the partial atp6 were 0.0%-1.3% between male and female parasites,and 0.0%-1.5% among isolates from different geographical origins.Phylogenetic analysis revealed that S.japonicum in Hunan province could not be clustered in accordance with related geographic origins.Sequence variation in apt6 gene among S.japonicum in Hunan province existed to a certain extent.An intensive study is worthy to analyze whether it was caused by the selection pressure of environment.